中国组织工程研究

中国组织工程研究 ›› 2024, Vol. 28 ›› Issue (1): 44-49.doi: 10.12307/2023.916

• 骨髓干细胞 bone marrow stem cells • 上一篇    下一篇

骨髓间充质干细胞来源外泌体调节大鼠肝细胞凋亡的机制

郑嵘炅，邓泽润，韩  丹，孙丽华   

  1. 新疆医科大学第一附属医院感染病·肝病中心，新疆维吾尔自治区乌鲁木齐市   830000
  • 收稿日期:2022-10-31 接受日期:2023-01-29 出版日期:2024-01-08 发布日期:2023-06-28
  • 通讯作者: 孙丽华，主任医师，新疆医科大学第一附属医院感染病·肝病中心，新疆维吾尔自治区乌鲁木齐市 830000
  • 作者简介:郑嵘炅，女，1981年生，新疆维吾尔自治区塔城市人，汉族，2018年新疆医科大学毕业，博士，副主任医师，主要从事感染性疾病及肝病的研究。
  • 基金资助:
    新疆维吾尔自治区自然科学基金项目(2020D01C243)，项目负责人：郑嵘炅

Mechanism underlying rat hepatocyte apoptosis regulated by exosomes derived from bone marrow mesenchymal stem cells

Zheng Rongjiong, Deng Zerun, Han Dan, Sun Lihua   

  1. Infection and Liver Disease Center of First Affiliated Hospital of Xinjiang Medical University, Urumqi 830000, Xinjiang Uygur Autonomous Region, China
  • Received:2022-10-31 Accepted:2023-01-29 Online:2024-01-08 Published:2023-06-28
  • Contact: Sun Lihua, Chief physician, Infection and Liver Disease Center of First Affiliated Hospital of Xinjiang Medical University, Urumqi 830000, Xinjiang Uygur Autonomous Region, China
  • About author:Zheng Rongjiong, MD, Associate chief physician, Infection and Liver Disease Center of First Affiliated Hospital of Xinjiang Medical University, Urumqi 830000, Xinjiang Uygur Autonomous Region, China
  • Supported by:
    Natural Science Foundation of Xinjiang Uygur Autonomous Region, No. 2020D01C243 (to ZRJ)

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摘要：


文题释义：

骨髓间充质干细胞：骨髓原始间充质干细胞是骨髓基质干细胞，是人们在哺乳动物的骨髓基质中发现的一种具有分化形成骨、软骨、脂肪、神经及成肌细胞等多种分化潜能的细胞亚群。它们对骨髓中的造血干细胞不仅有机械支持作用，还能分泌多种生长因子来支持组织细胞再生和血管形成。

外泌体：是指包含了复杂 RNA和蛋白质的小膜泡，直径为40-100 nm的盘状囊泡。多种细胞在正常及病理状态下均可分泌外泌体，其主要是细胞内溶酶体微粒内陷形成的多囊泡体，经多囊泡体外膜与细胞膜融合后释放到胞外基质中。


背景：骨髓间充质干细胞可释放大量外泌体，关于骨髓间充质干细胞来源外泌体对肝细胞凋亡的影响以及具体机制还没有完全阐明。
目的：探索骨髓间充质干细胞来源外泌体所携带的miR-21-5p对大鼠肝脏细胞凋亡的影响及其作用机制。
方法：分离大鼠骨髓间充质干细胞，将miR-21-5p NC或miR-21-5p inhibitor转染到骨髓间充质干细胞中，采用超速离心法提取外泌体，命名为(BMSCs+miR-21-5p NC)-Exos，(BMSCs+miR-21-5p inhibitor)-Exos，将外泌体与BRL大鼠肝细胞共培养，观察抑制 miR-21-5p表达后对大鼠肝细胞凋亡的影响。通过双荧光素酶报告基因检测验证外泌体中miR-21-5p和PIK3R1之间的靶向关系；TUNEL检测外泌体中miR-21-5p直接靶向PIK3R1激活PI3K/AKT信号通路对BRL大鼠肝细胞凋亡的影响。

结果与结论：①双荧光素酶报告系统证实，PI3KR1野生型载体与 miR-21-5p mimics共转染293T细胞时的荧光素酶活性显著低于PI3KR1突变型载体共转染组，表明miR-21-5p可靶向结合PIK3R1；②TUNEL检测结果显示：相比于(BMSCs+miR-21-5p NC)-Exos组，(BMSCs+miR-21-5p inhibitor)-Exos处理后BRL肝细胞凋亡率显著增加；与(BMSCs+miR-21-5p NC)-Exos组相比，加入AKT抑制剂LY294002之后，细胞凋亡率显著增加；③结果提示：外泌体可能通过miR-21-5p直接靶向PIK3R1激活PI3K/AKT信号通路抑制BRL大鼠肝细胞凋亡。

https://orcid.org/0000-0002-8611-6113 (郑嵘炅) 

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

关键词: 骨髓间充质干细胞, 外泌体, miR-21-5p, 肝细胞, 凋亡, PIK3R1

Abstract: BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) can release a large number of exosomes (Exos). The effect of Exos derived from BMSCs on hepatocyte apoptosis and the specific mechanism has not been fully clarified.
OBJECTIVE: To explore the effect of miR-21-5p carried by Exos derived from BMSCs on apoptosis of rat liver cells and its mechanism. 
METHODS: Rat BMSCs were isolated and miR-21-5p NC or miR-21-5p inhibitor was transfected into BMSCs. The Exos were extracted by ultracentrifugation and named (BMSCs+miR-21-5p NC)-Exos and (BMSCs+miR-21-5p inhibitor)-Exos. BMSCs-derived Exos were co-cultured with rat hepatocytes to observe the effect of inhibiting miR-21-5p expression on the apoptosis of rat hepatocytes. The targeting relationship between miR-21-5p and PIK3R1 was verified by double luciferase reporter gene detection. TUNEL was used to detect the effect of miR-21-5p directly targeting PIK3R1 in Exos to activate the PI3K/AKT signaling pathway on hepatocyte apoptosis in BRL rats.
RESULTS AND CONCLUSION: (1) The double luciferase reporting system confirmed that when PI3KR1 wild type vector and miR-21-5p mimics co-transfected 293T cells, the luciferase activity decreased significantly compared with the PI3KR1 mutant vector co-transfected group, indicating that miR-21-5p could target PIK3R1. (2) TUNEL test results showed that compared with (BMSCs+miR-21-5p NC)-Exos group, (BMSCs+miR-21-5p inhibitor)-Exos treatment significantly increased the apoptosis rate. Compared with the (BMSCs+miR-21-5p NC)-Exos group, after the addition of AKT inhibitor LY294002, the apoptosis rate was significantly increased. (3) The results indicate that Exos may inhibit the apoptosis of BRL rat hepatocytes through miR-21-5p, in which miR-21-5p directly targets PIK3R1 to activate PI3K/AKT signaling pathway. 

Key words: bone marrow mesenchymal stem cell, exosome, miR-21-5p, hepatocyte, apoptosis, PIK3R1

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